Identification of Host and Viral Determinants of Human Norovirus B cell Infection

人诺如病毒 B 细胞感染的宿主和病毒决定因素的鉴定

• 批准号: 10062847
• 负责人: Stephanie M Karst
• 金额: $ 47.78万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-12-01 至 2022-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10062847
• 关键词: Antiviral Agents; Attenuated Vaccines; B-Lymphocyte Subsets; B-Lymphocytes; Bile Acids; CRISPR/Cas technology; Cell Culture System; Cell Culture Techniques; Cell Cycle; Cells; Cessation of life; Child; Childhood; Clinic Visits; Clone Cells; Communities; Development; Diagnostic; Diarrhea; Disease; Disease Outbreaks; Drug Targeting; G1 Phase; Gastroenteritis; Generations; Genome; Genotype; Heparitin Sulfate; Hepatitis C virus; Human; In Vitro; Infection; Integration Host Factors; Interferons; Intervention; Intestines; Knock-out; Laboratories; Lead; Libraries; Mediating; Molecular; Mutation; Norovirus; Pharmaceutical Preparations; Phenotype; Poverty; Predisposition; Prevention strategy; Production; Property; Proteins; Publishing; Recording of previous events; Reporting; Research; Science; Serial Passage; Support System; Surface; Surveys; System; Testing; Text; Time; United States; Vaccines; Viral; Viral Antibodies; Viral Genome; Virus; Virus Replication; anti-hepatitis C; cell type; commensal bacteria; diagnostic platform; diarrheal disease; experimental study; fitness; foodborne illness; genome-wide; improved; in vivo; infected B cell; innovation; insight; novel; novel therapeutics; pandemic disease; prevent; response; reverse genetics; therapeutic development; tool; vaccine candidate; virus host interaction

SUMMARY Human noroviruses are a leading global cause of severe childhood diarrhea and gastroenteritis outbreaks. However, development of therapeutic and preventative strategies has been severely limited by the inability to propagate these viruses in culture. We recently discovered that noroviruses infect B cells in vitro and in vivo, and that they utilize commensal bacteria as a co-factor for infection. These novel findings led directly to our development of the first human norovirus cultivation system, a tool that will revolutionize norovirus research. We are now uniquely poised to identify host restriction and susceptibility factors influencing human norovirus infection of B cells as well as viral determinants of B cell infection. We hypothesize that this information will reveal key virus-host interactions that can guide the development of novel therapeutics, vaccine candidates, and enhanced diagnostic strategies. In Specific Aim 1, we will utilize complementary targeted and unbiased approaches to develop a comprehensive set of host factors that either restrict or promote human norovirus infection of B cells. In Specific Aim 2, a multi-pronged approach will be undertaken to identify viral determinants of in vitro replicative fitness. Strategies include serial passaging of virus in permissive cells and testing a diverse set of human norovirus genotypes for B cell infectivity. We will also construct a human norovirus reverse genetics system to test adaptive mutations for phenotypic effects. These studies will not only provide critical insight into host and viral determinants of B cell infection by human noroviruses but they should also culminate in the development of 2nd generation culture systems supporting robust infection of a broad panel of viral genotypes.

总结 人类诺如病毒是全球严重儿童腹泻和胃肠炎爆发的主要原因。 然而，治疗和预防策略的发展受到了严重限制， 在培养基中繁殖这些病毒。我们最近发现诺如病毒在体外和体内感染B细胞， 并且它们利用肠道细菌作为感染的辅助因子。这些新的发现直接导致了我们的 开发了第一个人类诺如病毒培养系统，这一工具将彻底改变诺如病毒研究。 我们现在正处于独特的准备，以确定主机限制和易感性因素影响人类 诺如病毒感染B细胞以及B细胞感染的病毒决定簇。我们假设这 这些信息将揭示关键的病毒-宿主相互作用，从而指导新疗法的开发， 候选疫苗和增强的诊断策略。在具体目标1中，我们将利用互补性 有针对性和公正的方法，以制定一套全面的东道国因素， 促进人诺如病毒感染B细胞。具体目标2将采取多管齐下的办法 来确定病毒在体外复制适应性的决定因素。策略包括病毒的连续传代 允许的细胞和测试一组不同的人诺如病毒基因型的B细胞感染性。我们还将 构建人诺如病毒反向遗传学系统以测试适应性突变的表型效应。这些 这些研究不仅提供了对人类感染B细胞的宿主和病毒决定因素的重要认识， 诺如病毒，但他们也应该在第二代培养系统的发展达到高潮， 广泛的病毒基因型的强烈感染。

项目成果

The major targets of acute norovirus infection are immune cells in the gut-associated lymphoid tissue.

• DOI: 10.1038/s41564-017-0057-7
• 发表时间: 2017-12
• 期刊: Nature microbiology
• 影响因子: 28.3
• 作者: Grau KR;Roth AN;Zhu S;Hernandez A;Colliou N;DiVita BB;Philip DT;Riffe C;Giasson B;Wallet SM;Mohamadzadeh M;Karst SM
• 通讯作者: Karst SM

Genome-scale CRISPR screens identify host factors that promote human coronavirus infection.

• DOI: 10.1186/s13073-022-01013-1
• 发表时间: 2022-01-27
• 期刊: Genome medicine
• 影响因子: 12.3
• 作者: Grodzki M;Bluhm AP;Schaefer M;Tagmount A;Russo M;Sobh A;Rafiee R;Vulpe CD;Karst SM;Norris MH
• 通讯作者: Norris MH